Caratterizzazione della microflora spontanea di un alimento fermentato vegano

Tra i prodotti vegani più richiesti vi sono i “formaggi” vegani, alimenti fermentati a base di frutta secca o ottenuti attraverso trattamenti su latte di mandorla e latte di soia, e successivamente fermentati. Nella mia attività ho caratterizzato un alimento fermentato vegano e studiato la successione microbica durante la fermentazione di un “formaggio” ottenuto partendo da anacardi e preparato in maniera artigianale. Oltre agli aspetti microbiologici, ho analizzato anche alcune caratteristiche fisico chimiche. Durante il processo di produzione gli anacardi vengono messi in ammollo per 8 ore a temperatura ambiente e, successivamente, i semi vengono scolati e risciacquati sotto acqua corrente. Gli anacardi vengono poi addizionati di acqua e microrganismi probiotici e tritati in un mixer fino al raggiungimento di una crema omogenea. A questo punto il prodotto viene lasciato riposare a temperatura ambiente per 48 ore durante le quali ha luogo la fermentazione e poi addizionato di ingredienti. Le indagini chimico fisiche effettuate hanno evidenziato che il pH si mostra già basso prima dell’inizio della fermentazione vera e propria e scende a 4.5 dopo 48 ore di riposo a causa dell’accumulo di acidi organici, ed in particolare di acido lattico e acetico che indicando un’attività fermentativa condotta dai batteri lattici. Le analisi microbiologiche hanno confermato che l’effettivo agente di fermentazione era costituito da questi batteri che sono stati identificati a livello molecolare. Le specie identificate due eterofermentanti (Weissella e Leuconostoc), presenti soprattutto nelle prime fasi della fermentazione, ed una omofermentante (Pediococcus), prende il sopravvento mano a mano che la fermentazione procede. Il lavoro svolto ha permesso di ottenere alcune importanti informazioni per la produzione industriale di un “formaggio” vegano fermentato. Il processo studiato presenta numerosi punti di rischio che devono essere presi in considerazione prima di poter giungere alla messa a punto di un prodotto definitivo.

Metabolismo di oligosaccaridi prebiotici in Bifidobacterium per il potenziale sviluppo di nuovi prodotti alimentari funzionali

The growth and the metabolism of Bifidobacterium adolescentis MB 239 fermenting GOS, lactose, galactose, and glucose were investigated. An unstructerd unsegregated model for growth of B. adolescentis MB 239 in batch cultures was developed and kinetic parameters were calculated with a Matlab algorithm. Galactose was the best carbon source; lactose and GOS led to lower growth rate and cellular yield, but glucose was the poorest carbon source. Lactate, acetate and ethanol yields allowed calculation of the carbon fluxes toward fermentation products. Similar distribution between 3- and 2-carbon products was observed on all the carbohydrates (45 and 55%, respectively), but ethanol production was higher on glucose than on GOS, lactose and galactose, in decreasing order. Based on the stoichiometry of the fructose 6-phosphate shunt and on the carbon distribution among the products, ATP yield was calculated on the different carbohydrates. ATP yield was the highest on galactose, while it was 5, 8, and 25% lower on lactose, GOS, and glucose, respectively. Therefore, a correspondance among ethanol production, low ATP yields, and low biomass production was established demonstrating that carbohydrate preferences may result from different sorting of carbon fluxes through the fermentative pathway. During GOS fermentation, stringent selectivity based on the degree of polymerization was exhibited, since lactose and the trisaccharide were first to be consumed, and a delay was observed until longer oligosaccharides were utilized. Throughout the growth on both lactose and GOS, galactose accumulated in the cultural broth, suggesting that β-(1-4) galactosides can be hydrolysed before they are taken up. The physiology of Bifidobacterium adolescentis MB 239 toward xylooligosaccharides (XOS) was also studied and our attention was focused on an extracellular glycosyl-hydrolase (β-Xylosidase) expressed by a culture of B. adolescentis grown on XOS as sole carbon source. The extracellular enzyme was purified from the the supernatant, which was dialyzed and concentrated by ultrafiltration. A two steps purification protocol was developed: the sample was loaded on a Mono-Q anion exchange chromatography and then, the active fractions were pooled and β-Xylosidase was purified by gel filtration chromatography on a Superdex-75. The enzyme was characterized in many aspects. β- Xylosidase was an homo-tetramer of 160 kDa as native molecular mass; it was a termostable enzyme with an optimum of temperature at 53 °C and an optimum of pH of 6.0. The kinetics parameter were calculated: km = 4.36 mM, Vmax = 0.93 mM/min. The substrate specificity with different di-, oligo- and polysaccharides was tested. The reactions were carried out overnight at pH 7 and at the optimum of temperature and the carbohydrates hydrolysis were analyzed by thin layer chromatography (TLC). Only glycosyl-hydrolase activities on XOS and on xylan were detected, whereas sucrose, lactose, cellobiose, maltose and raffinose were not hydrolyzed. It’s clearly shown that β-Xylosidase activity was higher than the Xylanase one. These studies on the carbohydrate preference of a strain of Bifidobacterium underlined the importance of the affinity between probiotics and prebiotics. On the basis of this concept, together with Barilla G&R f.lli SpA, we studied the possibility to develop a functional food containing a synbiotic. Three probiotic strains Lactobacillus plantarum BAR 10, Streptococcus thermophilus BAR 20, and Bifidobacterium lactis BAR 30 were studied to assess their suitability for utilization in synbiotic products on the basis of antioxidative activity, glutathione production, acid and bile tolerance, carbohydrates fermentation and viability in food matrices. Bile and human gastric juice resistance was tested in vitro to estimate the transit tolerance in the upper gastrointestinal tract. B. lactis and L. plantarum were more acid tolerant than S. thermophilus. All the strains resisted to bile. The growth kinetics on 13 prebiotic carbohydrates were determined. Galactooligosaccharides and fructo-oligosaccharides were successfully utilized by all the strains and could be considered the most appropriate prebiotics to be used in effective synbiotic formulations. The vitality of the three strains inoculated in different food matrices and maintained at room temperature was studied. The best survival of Lactobacillus plantarum BAR 10, Streptococcus thermophilus BAR 20, and Bifidobacterium lactis BAR 30 was found in food chocolate matrices. Then an in vivo clinical trial was carried out for 20 healthy volunteers. The increase in faecal bifidobacteria and lactobacilli populations and the efficacy of the pre-prototype was promising for the future develop of potential commercial products.

Prove di ozonolisi sui fanghi biologici prodotti da impianti di depurazione delle acque reflue urbane

La tecnica di ozonolisi viene applicata ai fanghi biologici derivanti da impianti di depurazione acque reflue urbane, e consiste nell'ottenere, grazie all'ozono, una minor massa fangosa da smaltire e una miglior trattabilità del fango residue. In questo elaborato si prendono in esame le sperimentazioni effettuate a Marina di Ravenna e si estraggono le prime conclusioni gestionali, economiche e ambientali sull'applicabilità del metodo a questo tipo di fango.

Lo sviluppo nuovo prodotti nel settore auto motive: analisi dei processi organizzativi attraverso lo studio di un caso

Mappatura dei processo organizzativi, della struttura organizzativa e dei sistemi informativi di supporto. analisi di alcune problematiche riscontrate

Messa a punto di procedure per la semi-sintesi di composti ad attività antitumorale partendo da prodotti naturali. Studi di struttura-attività

Camptothecin, (CPT) is a pentacyclic alkaloid isolated for the first time from the Chinese tree Camptotheca acuminata, and which has soon attracted the attention of medicinal chemists and pharmacologists due to its promising anti-cancer activity against the most aggressive histo-types. So far, most of the synthesized camptothecin analogues are A and B ring modified compounds, which have been prepared via synthetic or semi-synthetic routes. To the best of our knowledge, a very limited number of C, D, or E ring modified analogues of CPT have been reported; moreover, the few derivatives known from the literature showed a reduced or no biological activity. This dissertation presents synthetic studies on camptothecin new derivatives along with the development of a new and general semi-synthetic methodology to obtain a large variety of analogues. We report here the semi-synthesis of a new family of 5-substituted CPT's, along with their biological activity evaluation, which will be compared with reference compounds. The use of carrier-linked prodrugs has emerged as a useful strategy to overcome some of the drawbacks related with the use of the parent drug, such as low solubility, membrane permeability properties, low oral absorption, instability, toxicity, and nontargeting. Herein we report CPT-prodrugs synthesized via ring opening of the lactone moiety as 17-O-acyl camptothecin tripartate conjugates, which bear a polyamine side chain with different architectures, as the carriers. Moreover, we found that the replacement of the oxygen atom with sulphur on the piridone D-ring, dramatically improves the potency of the novel 16a-thio-camptothecin derivatives, opening new possibilities in the modelling of this class of compounds.